应用间接ELISA检测鸡传染性鼻气管炎病毒抗体

Indirect ELISA for Detection of Avain Rhinotracheitis Virus in Chickens

采用增殖的鸡传染性鼻气管炎病毒 (ARTV) NL7784株 ,经 PEG- 2 0 0 0 0浓缩液提纯抗原和纯化羊抗鸡 Ig G抗体 ,建立了检测 ARTV抗体的间接酶联免疫吸附试验。该试验的最佳反应条件为 :包被液为 p H9.6的 0 .0 5 mol/ L碳酸盐缓冲液 ,抗原最佳稀释度为 1∶ 16 0 ,以含 6 %犊牛血清 (CS)的磷酸盐缓冲液作为封闭液 ,稀释液用含 10 %CS、0 .5 %Tween- 2 0的磷酸盐缓冲液 ,抗原与血清结合的最佳反应时间为 30 min;酶标抗体的最佳稀释倍数为 1∶ 10 0 0 ,酶联反应的最佳时间为 30 min,底物的最佳反应时间为 15 min。本方法具有良好的特异性、稳定性、可重复性和较高的敏感性。

Indirect-ELISA for estimation of avian rhinotracheitis virus were developed by using purified ARTV antigen and antichicken conjugated with horseradish peroxidas(HRP),the whole procedure was optimized.Polyoxyl(PEG-20000) dense were coated with purified ARTV antigen at the amount of 0.05 mol/L sodium bicarbonate buffer(pH9.6) per well.The working concentration of HRP-labelled sheep antichicken IgG and that of serum samples respectively was 1∶160,6% CS seal,0.5% Tween-20 PBST dilution in detection of IgG.These ELISA systems were proved to be sensitive and specific.